The engineered E. coli is the first consolidated advanced biofuels production and cellulosic bioprocessing path. The breakthrough enables the production of advanced hydrocarbon fuels and chemicals in a single fermentation process that does not require additional chemical transformations.

E coli Secreting Bio Diesel LS9 Click image for more info.

This week LS9 will announce the planned location of a demonstration facility in the U.S. to convert sugar cane into biodiesel using an existing organism from previous work. Stephen del Cardayre, the vice president of research and development at the company said the plant, which will use an existing engineered microorganism, will open this summer and pave the way for large-scale manufacturing and sales in 2012.

The new E. coli is a step toward lowering the cost of making biodiesel from wood chips, corn stover, and other residual agricultural products. The added enzymes made from the new genetic modifications greatly expand the feedstock choices including the still rare switchgrass and miscanthus. These types of cellulosic feedstocks are typically much harder to convert into fuel through fermentation than sugar cane or corn, but offer the potential of more complete CO2 cycling through the atmosphere.

The researcher’s design imports genes that allow E. coli to secrete enzymes that break down the tough material that makes up the bulk of plants – cellulose, specifically hemicellulose – and produce the sugar needed to feed the process. Chemical engineer Jay Keasling of the University of California, Berkeley says, “The organism can produce the fuel from a very inexpensive sugar supply, namely cellulosic biomass.”

Keasling explains then, “We incorporated genes that enabled production of biodiesel – esters [organic compounds] of fatty acids and ethanol – directly. The fuel that is produced by our E. coli can be used directly as biodiesel. In contrast, fats or oils from plants must be chemically esterified before they can be used.” This eliminates one process and it seems, no glycerin is produced.

The E. coli directly secretes the resulting biodiesel, which then floats to the top of a fermentation vat, so there is neither the necessity for distillation or other purification processes nor the need, as in biodiesel from algae as developed so far, to break the cell to get the oil out.

Keasling and his team cloned genes from Clostridium stercorarium and Bacteroides ovatus, bacteria species that live in soil and the guts of plant-eating animals that produce enzymes for breaking down the cellulose. The team then used the extra bits of genetic code in the form of short amino acid sequences that instruct the altered E. coli to secrete the bacterial enzyme out into the feedstock, which breaks down the plant cellulose, turning it into sugar; then the E. coli in turn transforms that sugar into biodiesel.

Keasling points out the new E. coli are not the most efficient producer of biofuel, “We are at about 10 percent of the theoretical maximum yield from sugar. We would like to be at 80 to 90 percent to make this commercially viable. Furthermore, we would need a large-scale production process,” to allow mass production of microbial fuel.

LS9 isn’t alone at this either, Gevo and Keasling’s own founded Amyris Biotechnologies, are working on making fuel from microbes commercially viable.

Keasling explains the idea in this case is to produce a batch of biofuel from a single colony through E. coli’s natural ability to proliferate and, after producing the fuel, dispose of the E. coli and start anew with a fresh colony, “This minimizes the mutations that might arise if one continually subcultured the microbe.” The idea is also to engineer the new organism, deleting key metabolic pathways, such that it would never survive in the wild in order to prevent escapes with unintended environmental impacts, among other dangers.

Even at just 10% of the carbohydrates available going to a product, the first path for a single step process seems to be in hand. 80 or 90% is a ways off. The competition may well find that a two-step process yields more net income. $75 oil makes a lot of possibilities viable.

If – and it’s a major question, the process can utilize crops from more marginal land and offer a residue worth returning to the soil instead of losing the major trace elements such as potassium and phosphorus, growing fuels in the higher carbon molecules could really change the function of fuel markets and agriculture.

What we can envision is the day when farmers have silos where you drop silage into the top and remove digested fertilizer from the bottom – with a steady trickle of biodiesel coming out to the holding tank.

When farmers have a self contained biodiesel system – the american food supply will be protected from oil shocks.

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